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WO2019089365A1 - Nouvelles formes solides de squalamine et procédés pour les produire - Google Patents

Nouvelles formes solides de squalamine et procédés pour les produire Download PDF

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Publication number
WO2019089365A1
WO2019089365A1 PCT/US2018/057650 US2018057650W WO2019089365A1 WO 2019089365 A1 WO2019089365 A1 WO 2019089365A1 US 2018057650 W US2018057650 W US 2018057650W WO 2019089365 A1 WO2019089365 A1 WO 2019089365A1
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Prior art keywords
squalamine
solid form
squalamine phosphate
phosphate
theta
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William A. Kinney
Michael Zasloff
Anil Kumar
Amanuel Z. ZELELLOW
Dalian Zhao
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Enterin Laboratories Inc
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Enterin Laboratories Inc
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Priority to EP18874567.3A priority Critical patent/EP3704132A4/fr
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J41/00—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring
    • C07J41/0005—Normal steroids containing one or more nitrogen atoms not belonging to a hetero ring the nitrogen atom being directly linked to the cyclopenta(a)hydro phenanthrene skeleton
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/08—Antibacterial agents for leprosy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J31/00—Normal steroids containing one or more sulfur atoms not belonging to a hetero ring
    • C07J31/006—Normal steroids containing one or more sulfur atoms not belonging to a hetero ring not covered by C07J31/003

Definitions

  • the present technology relates generally to solid forms of squalamine phosphate.
  • squalamine presented a structure never before seen in nature, that being a bile acid coupled to a polyamine (spermidine); i.e. , a steroid chemically linked to a polyamine.
  • spermidine polyamine
  • the chemical structure of squalamine also known as 3 beta-N-l-(N-[3-(4-aminobutyl)]- 1,3- diaminopropane)-7 alpha,24 zeta-dihydroxy-5 alpha-cholestane 24-sulfate, has been determined by fast atom bombardment mass spectroscopy and NMR.
  • Squalamine is a cationic steroid characterized by a condensation of an anionic bile salt intermediate with spermidine.
  • Squalamine is a cationic amphipathic substance exhibiting an affinity for membranes composed of anionic phospholipids (Selinsky et al., "The aminosterol antibiotic squalamine permeabilizes large unilamellar phospholipid vesicles,” Biochim. Biophys. Acta., 1370(2): 218-34 (1998); Selinsky et al, "Squalamine is not a proton ionophore," Biochim.
  • squalamine is believed to exert antimicrobial action by interacting electrostatically with the membranes of target microorganisms, which generally display anionic phospholipids on the membrane surface exposed to the environment,
  • Genaera Corporation discontinued trials for the use of squalamine in treating cancer in 2007.
  • Squalamine is also marketed under the brand name SqualamaxTM as a dietary supplement, though it has not been approved as a drug in this form and thus cannot make therapeutic claims.
  • SqualamaxTM is an unfractionated extract of shark liver, containing innumerable uncharacterized substances in addition to squalamine, and squalamine is present in SqualamaxTM at less than 0.01% of the total weight of the extract. "Cyber Warning Letter", Center for Drug Evaluation and Research (2002-05-06),
  • the dietary supplement form of squalamine is not pharmaceutical grade squalamine, as pharmaceutical grade squalamine requires significantly greater manufacturing efforts.
  • an isolated solid form of squalamine phosphate designated as Form 1.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 15.7° and about 23.7°.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 1 1 .7°, about 15.7°, about 19.7°, and about 23.7°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 8.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C. In some embodiments, the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9. In some embodiments, the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 10. In some embodiments, the solid form is substantially purified. In some embodiments, the solid form has a water content of less than about 8%. In some embodiments, the solid form has a water content of about 6-8%.
  • a solid form of squalamine phosphate having an X- ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 15.7° and at about 23.7°.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 8.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C.
  • DSC differential scanning calorimetry
  • the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9. In some embodiments, the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 10. In some embodiments, the solid form is substantially purified. In some embodiments, the solid form has a water content of less than about 8%. In some embodiments, the solid form has a water content of about 6-8%.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • a crystalline squalamine phosphate characterized by an X-ray powder diffraction pattern comprising the following peaks: about 15.7° 2-theta and about 23.7° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • an isolated squalamine phosphate solid form designated as Form 2.
  • the solid form has an X-ray powder diffraction pattern comprising a peak, in terms of 2-theta, at about 1 .2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern comprising a peak, in terms of 2-theta, at about 1 1.4°, at about 15.2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 15.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C. In some embodiments, the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 16. In some embodiments, the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 17. In some embodiments, the solid form is substantially purified. In some embodiments, the solid form has a water content of about 9-12%. In some embodiments, the solid form has a water content of more than about 9%.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • a solid form of squalamine phosphate having an X- ray powder diffraction pattern comprising a peak, in terms of 2-theta, at about 15.2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C. In some embodiments, the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 16. In some embodiments, the solid form has a thermo gravimetric analysis (TGA) substantially as shown in FIG. 17. In some embodiments, the solid form is substantially purified. In some embodiments, the solid form has a water content of about 9-12%. In some embodiments, the solid form has a water content of more than about 9%.
  • a crystalline squalamine phosphate characterized by an X-ray powder diffraction pattern comprising the following peaks: at about 1 1.4° 2-theta, at about 15.2° 2-theta and at about 22.9° 2-theta, as determined on a diffractometer using Cu-Ka radiation at a wavelength of 1.5406 A.
  • composition comprising the squalamine phosphate solid form of Form 1 or Form 2, and a pharmaceutically acceptable carrier.
  • compositions comprising squalamine phosphate, wherein the squalamine phosphate in the composition comprises the squalamine phosphate solid form of Form 1 or Form 2 in an amount of at least about 90% by weight.
  • compositions consisting essentially of the squalamine phosphate solid form of Form 1 or Form 2.
  • composition comprising the squalamine phosphate solid form of Form 1 , the squalamine phosphate solid form of Form 2, and a pharmaceutically acceptable carrier.
  • compositions comprising squalamine phosphate, wherein the squalamine phosphate in the composition comprises the squalamine phosphate solid form of Form 1 and the squalamine phosphate solid form of Form 2 in a total amount of at least about 90% by weight.
  • compositions consisting essentially of the squalamine phosphate solid form of Form 1 and the squalamine phosphate solid form of Form 2.
  • methods for treating a subject in need having a condition susceptible to treatment with an aminosterol comprising administering to the subject a therapeutically effective amount of the squalamine phosphate solid form of Form 1 and/or Form 2, or a pharmaceutical composition comprising the squalamine phosphate solid form of Form 1 and/or Form 2.
  • provided herein are methods for treating a subject in need having a condition susceptible to treatment with an aminosterol, comprising administering to the subject a therapeutically effective amount of a composition comprising or consisting essentially of a squalamine phosphate solid form of Form 1 and/or Form 2, and one or more
  • the subject in need has a condition selected from the group consisting of viral infections, antimicrobial infections, Gram-negative and Gram-positive bacterial infections, Mycobacteria infections, fungal infections, protozoan infections, disease states known to be associated with pathological neovascularization, such as cancer, vascular disorders of the eye, including macular degeneration, such as age-related macular degeneration, retinopathy of prematurity, corneal neovascularization, diabetic retinopathy, weight loss or weight management, dose-dependent weight loss; diseases, including viral infections, where sodium-hydrogen exchanger ("NHE-3") plays a critical role, treatment of fibrodysplasia ossificans progressiva, disorders of neovascularization, and sleep disorders.
  • NHE-3 sodium-hydrogen exchanger
  • the viral infection is caused by a virus selected from the group consisting of Yellow Fever, Cytomegalovirus, Eastern Equine Encephalitis virus, Hepatitis B virus, Hepatitis Delta virus, Dengue virus, and Human Immunodeficiency virus.
  • the condition to be treated is a viral infection caused by a virus selected from the group consisting of "African Swine Fever Viruses," Arbovirus, Adenoviridae, Arenaviridae, Arterivirus,
  • Paramyxoviridae Morbillivirus, Rhabdoviridae), Myoviridae, Orthomyxoviridae (e.g., Influenza A, Influenza B, and parainfluenza), Papiloma virus, Papovaviridae, Paramyxoviridae, Prions, Parvoviridae, Phycodnaviridae, Picomaviridae (e.g., Rhinovirus, Poliovirus), Poxviridae (such as Smallpox or Vaccinia), Potyviridae, Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-I, HTLV- II, Lentivirus), Rhabdoviridae, Tectiviridae, Togaviridae (e.g., Rubivirus), herpes, pox, papilloma, corona, influenza, hepatitis, sendai, Sindbis, vaccinia viruses, west
  • the condition to be treated is selected from the group consisting of AIDS, viral meningitis, Dengue, EBV, hepatitis, a chronic disease suspected to be of viral origin, multiple sclerosis, Type I diabetes, Type II diabetes, atherosclerosis, cardiomyopathies, awaski disease, aplastic anemia, and any combination thereof.
  • the condition to be treated is a neurological disorder selected from the group consisting of Parkinson's disease, autism, multiple system atrophy, depression, Alzheimer's disease, Huntington's Disease, schizophrenia, multiple sclerosis, and degenerative processes associated with aging, autonomic system instability, circadian rhythm disruption, delays in sleep onset, fragmentation of sleep, reduced REM sleep, reduced total sleep time, REM-behavior disorder, sleep breathing disorder including snoring and sleep apnea, hallucinations, narcolepsy, and day-time sleepiness.
  • the condition to be treated is a gastrointestinal disorder selected from the group consisting of constipation, inflammatory bowel disease, and irritable bowel syndrome.
  • the method further comprises administering an additional active agent, wherein the additional active agent is administered via a method selected from the group consisting of (a)
  • the subject is human.
  • the subject in need has a condition selected from the group consisting of disorders of gastrointestinal motility, such as chronic idiopathic constipation, opioid induced constipation, irritable bowel syndrome, and inflammatory bowel disease; diabetes mellitus and diabetic neuropathy; disorders of the nervous system that could benefit from neuro-protection, such as Parkinson's Disease, Alzheimer's disease, Huntington's Disease, acute traumatic injury to the central nervous system, including the spinal cord, stroke, acute head and/or spine injury, degenerative processes associated with aging, including memory loss ("dementia of aging"), cerebral palsy, epilepsy, peripheral sensory neuropathy, and multiple sclerosis; autism; sleep disorders; schizophrenia; depression; and autonomic system lability.
  • disorders of gastrointestinal motility such as chronic idiopathic constipation, opioid induced constipation, irritable bowel syndrome, and inflammatory bowel disease
  • diabetes mellitus and diabetic neuropathy disorders of the nervous system that could benefit from neuro-protection, such as Parkinson'
  • processes for preparing the squalamine phosphate solid form of Form 1 comprising: (a) combining a solution of squalamine lactate with an aqueous solution of sodium phosphate dibasic and sodium phosphate monobasic to form a combined mixture; (b) heating the combined mixture before allowing the combined mixture to cool; and (c) isolating the combined mixture to isolate the squalamine phosphate solid form.
  • the combining step is performed at a temperature of about 55 °C to about 70 °C. In some embodiments, the combining step is performed at 60 ⁇ 5 °C.
  • the solution of squalamine lactate is a methanol solution of squalamine lactate.
  • the heating step is performed at 70 ⁇ 5 °C.
  • the process further comprises stirring the combined mixture after cooling but before the isolating step.
  • the isolating step comprises filtering the combined mixture and washing filtered solids with acetone.
  • processes for preparing the squalamine phosphate solid form of Form 2 comprising: (a) dissolving squalamine lactate in base, water, and alcohol to form a first solution; (b) heating the first solution to a first elevated temperature, wherein the first elevated temperature is greater than 25 °C; (c) adding a first amount of phosphoric acid (H3PO4) to the first solution to form a second solution; (d) heating the second solution to a second elevated temperature higher than the first elevated temperature; (e) adding a second amount of H 3 PO 4 to the second solution; (f) obtaining a slurry; and (h) isolating the squalamine phosphate solid form.
  • H3PO4 phosphoric acid
  • the base is sodium hydroxide.
  • the alcohol is ethanol.
  • the first elevated temperature is at least about 35 °C.
  • the second elevated temperature is at least about 45 °C.
  • the step of obtaining the slurry comprises seeding the mixture.
  • the step of obtaining the slurry comprises adding a third amount of H3PO4 to the second solution.
  • the step of obtaining the slurry comprises cooling the second solution.
  • the process further comprises aging the slurry prior to the isolating step.
  • the isolating step comprises filtering the slurry and washing filtered solids with acetone.
  • FIG. 1 shows X-ray powder diffraction (XRPD) patterns of squalamine phosphate samples of (a) dry product of initial preparation (Example 2), (b) final wet cake of initial preparation (Example 2), and (c) Form 1 (from Example 5).
  • XRPD X-ray powder diffraction
  • FIG. 2 shows XRPD patterns of squalamine phosphate samples of (a) Form 1 (from Example 5), and (b) end solids from crystallinity upgrade (Form 2, Example 2).
  • FIG. 3 shows results from thermogravimetric analysis (TGA) of squalamine phosphate, Form 2 (from Example 2).
  • FIG. 4 shows results from differential scanning calorimetry (DSC) of squalamine phosphate, Form 2 (from Example 2).
  • FIG. 5 shows XRPD patterns of squalamine phosphate samples of (a) Form 1 from Example 5, (b) Form 1 material (from Example 5) used as seed in Example 4, (c) Form 2 from Example 2 (after crystallinity upgrade), (d) wet cake (Form 2) of Example 4.
  • FIG. 6 shows results from TGA of squalamine phosphate, Form 2 (from Example 4).
  • FIG. 7 shows results from DSC of squalamine phosphate, Form 2 (from Example 4).
  • FIG. 8 shows an XRPD pattern of squalamine phosphate, Form 1.
  • FIG. 9 shows results from DSC of squalamine phosphate, Form 1.
  • FIG. 10 shows results from TGA of squalamine phosphate, Form 1.
  • FIG. 11 shows XRPD patterns of squalamine phosphate samples of (a) Form 1
  • Example 5 (Example 5), (b) after water-acetone wash of Form 2, (c) after water-ethanol wash of Form 2, and (d) Form 2 (Example 2, after crystallinity upgrade).
  • the HiO-acetone and H 2 0-EtOH washed samples match Form 1 (i.e., Form 2 converted to Form 1).
  • FIG. 12 shows XRPD patterns of squalamine phosphate samples of (a) Form 1
  • FIG. 13 shows XRPD patterns of squalamine phosphate samples of (a) Form 1
  • FIG. 14 shows XRPD patterns of squalamine phosphate samples of (a) Form 1
  • FIG. 15 shows an XRPD pattern of squalamine phosphate, Form 2.
  • FIG. 16 shows results from DSC of squalamine phosphate, Form 2.
  • FIG. 17 shows results from TGA of squalamine phosphate, Form 2.
  • the present technology relates to solid forms of squalamine phosphate, methods of preparing the solid forms, compositions comprising one or more of the solid forms of squalamine phosphate, and methods of use thereof.
  • the solid forms include crystalline solid forms.
  • squalamine has been explored as a pharmaceutical agent useful in treating a wide variety of conditions.
  • one of the challenges with methods of treatment using squalamine is identifying a repeatable source for squalamine. Isolation of the compound from its naturally occurring source, e.g., the dogfish shark, is not feasible for large scale use of squalamine.
  • synthetic methods of making squalamine have to date been complex, and therefore expensive, due to the structural complexity of the compound. Described herein are methods of making novel crystalline forms of squalamine that can be easily isolated and are suitable for oral and topical formulation. These new methods of chemical synthesis and isolation result in effective and novel forms of squalamine.
  • Squalamine phosphate which has limited solubility at neutral pH, has reduced bitterness compared with an equivalent concentration of the fully soluble salt.
  • the free concentration of the squalamine ion (the molecule recognized by the taste receptors) in the presence of phosphate is many orders of magnitude lower than it would be in the absence of phosphate, thereby reducing the effective concentration bathing the taste receptors on the tongue.
  • an isolated solid form of squalamine phosphate designated as Form 1.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 15.7° and at about 23.7°.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 8.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C. In some embodiments, the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9. In some embodiments, the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 10.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • a solid form having an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 15.7° and at about 23.7°.
  • the solid form has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 8.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C.
  • the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9.
  • the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 10.
  • a crystalline squalamine phosphate characterized by an X-ray powder diffraction pattern comprising the following peaks: about 15.7° 2-theta and about 23.7° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • the crystalline squalamine phosphate has an X-ray powder diffraction pattern comprising peaks, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°.
  • the crystalline squalamine phosphate has an X-ray powder diffraction pattern substantially as shown in FIG. 8.
  • the crystalline squalamine phosphate has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C. In some embodiments, the crystalline squalamine phosphate has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9. In some embodiments, the crystalline squalamine phosphate has a
  • thermogravimetric analysis substantially as shown in FIG. 10.
  • the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 1 5.7° 2-theta and about 23.7° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A; and (ii) a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C.
  • the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 11.7°, about 15.7°, about 19.7°, and about 23.7°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 15.7° 2-theta and about 23.7° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A; (ii) a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 96.6 °C; and (iii) has a thermo gravimetric analysis (TGA) substantially as shown in FIG. 10.
  • DSC differential scanning calorimetry
  • TGA thermo gravimetric analysis
  • the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 15.7° 2-theta and about 23.7° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A; (ii) a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 9; and (iii) has a thermogravimetric analysis (TGA) substantially as shown in FIG. 10.
  • the X-ray powder diffraction comprises a peak, in terms of 2- theta, at about 1 1.7°, about 15.7°, about 19.7°, and about 23.7°, as determined on a
  • the squalamine phosphate solid form of Form 1 may have a water content of less than or equal to about 8%. In some embodiments, the squalamine phosphate solid form of Form 1 has a water content of less than or equal to about 7%.
  • the squalamine phosphate solid form of Form 1 has a water content of less than or equal to about 7.5%, less than or equal to about 7%, less than or equal to about 6.5%, less than or equal to about 6%, less than or equal to about 5.5%, less than or equal to about 5%, less than or equal to about 4.5%, less than or equal to about 4%, less than or equal to about 3.5%, less than or equal to about 3%, less than or equal to about 2.5%, less than or equal to about 2%, less than or equal to about 1.5%, or less than or equal to about 1 .5%, including increments between any of these values.
  • the squalamine phosphate solid form of Form 1 may have a water content of about 6% to about 8%, or about 6% to about 7%, or about 7% to about 8%.
  • an isolated squalamine phosphate solid form designated as Form 2.
  • the solid form has an X-ray powder diffraction pattern comprising a peak, in terms of 2-theta, at about 15.2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern comprising a peak, in terms of 2-theta, at about 11.4°, at about 15.2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 15. In some
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • a solid form having an X-ray powder diffraction pattern comprising a eak, in terms of 2-theta, at about 15.2° and at about 22.9°.
  • the solid form has an X-ray powder diffraction pattern substantially as shown in FIG. 15.
  • the solid form has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C.
  • the solid form has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 16.
  • the solid form has a thermogravimetric analysis (TGA) substantially as shown in FIG. 17.
  • a crystalline squalamine phosphate characterized by an X-ray powder diffraction pattern comprising the following peaks: about 15.2° 2-theta and at about 22.9° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 ⁇ .
  • the crystalline squalamine phosphate has an X-ray powder diffraction pattern substantially as shown in FIG, 15.
  • the crystalline squalamine phosphate has a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C.
  • the crystalline squalamine phosphate has a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG. 16. In some embodiments, the crystalline squalamine phosphate has a thermogravimetric analysis (TGA) substantially as shown in FIG. 17. In some embodiments, the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 1 1.4°, about 15.2°, and about 22.9°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 15.2° 2-theta and at about 22.9° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A; and (ii) a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C.
  • the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 1 1.4°, about 15.2°, and about 22.9°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 15.2° 2-theta and at about 22.9° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406A; (ii) a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak at about 90.9 °C; and (iii) a thermogravimetric analysis (TGA) substantially as shown in FIG. 17.
  • DSC differential scanning calorimetry
  • TGA thermogravimetric analysis
  • the X-ray powder diffraction comprises a peak, in terms of 2-theta, at about 1 1.4°, about 15.2°, and about 22.9°, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406 A.
  • a crystalline squalamine phosphate characterized by (i) an X-ray powder diffraction pattern comprising the following peaks: about 15.2° 2-theta and at about 22.9° 2-theta, as determined on a diffractometer using Cu- ⁇ radiation at a wavelength of 1.5406A; (ii) a differential scanning calorimetry (DSC) thermogram substantially as shown in FIG.
  • the X-ray powder diffraction comprises a peak, in terms of 2- theta, at about 1 1.4°, about 15.2°, and about 22.9°, as determined on a diffractometer using Cu- Ka radiation at a wavelength of 1.5406 A.
  • the squalamine phosphate solid form of Form 2 may have a water content of equal to or greater than about 9%. This includes a water content of about 10%, 1 1 %, or 12%, including increments therein. In some embodiments, the squalamine phosphate solid form of Form 2 has a water content of more than about 10%.
  • the squalamine phosphate solid form of Form 2 has a water content equal to or greater than about 9%, equal to or greater than about 9.5%, equal to or greater than about 10%, equal to or greater than about 10.5%, equal to or greater than about 1 1%, equal to or greater than about 1 1.5%, equal to or greater than about 12%, equal to or greater than about 12.5%, equal to or greater than about 13%, equal to or greater than about 13.5%, equal to or greater than about 14%, equal to or greater than about 14.5%, or up to about 15%, including any amount in-between these values.
  • the squalamine phosphate solid form of Form 2 may have a water content of about 9% up to about 12%. This includes a water content of about 9%, about 10%, about 1 1%, or about 12%, including increments therein. In some embodiments, the squalamine phosphate solid form of Form 2 has a water content of about 9% to about 1 1%, about 9% to about 10%, about 10% to about 1 1%, about 10% to about 12%, or about 11% to about 12%.
  • compositions comprising Form 1 and/or Form 2 of squalamine phosphate.
  • compositions comprising the squalamine phosphate solid form of Form 1 , and one or more pharmaceutically acceptable carriers.
  • pharmaceutical compositions comprising the squalamine phosphate solid form of Form 1, and a pharmaceutically acceptable carrier.
  • compositions comprising the squalamine phosphate solid form of Form 2, and one or more pharmaceutically acceptable carriers.
  • pharmaceutical compositions comprising the squalamine phosphate solid form of Form 2, and a pharmaceutically acceptable carrier.
  • compositions comprising squalamine phosphate, wherein the squalamine phosphate in the composition comprises the squalamine phosphate solid form of Form 1 and/or Form 2 in an amount of at least about 88% by weight of the squalamine phosphate in the composition. This includes an amount of about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 9%7, about 98%, about 99%, or more, by weight of the squalamine phosphate in the composition, including increments therein.
  • a squalamine phosphate composition While it is possible for a squalamine phosphate composition to be administered alone, it is preferable to present it as a pharmaceutical formulation, together with one or more acceptable carriers.
  • the carrier(s) must be "acceptable” in the sense of being compatible with Form 1 and/or Form 2 of squalamine phosphate and not deleterious to the recipients thereof.
  • the formulations are prepared by contacting Form 1 and/or Form 2 of squalamine phosphate, uniformly and intimately with liquid carriers or finely divided solid carriers or both. Then, if necessary, the product is shaped into the desired formulation.
  • Nonaqueous vehicles such as fixed oils and ethyl oleate are also useful herein, as well as liposomes.
  • the carrier suitably comprises minor amounts of additives such as substances that enhance isotonicity and chemical stability.
  • additives such as substances that enhance isotonicity and chemical stability.
  • Such materials are non-toxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or their salts; antioxidants such as ascorbic acid; low molecular weight (less than about ten residues) polypeptides, e.g., polyarginine or tripeptides; proteins, such as gelatin, serum albumin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids, such as glycine, glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, manose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbi
  • Form 1 and/or Form 2 of squalamine phosphate can be formulated as aerosols using standard procedures.
  • aerosol includes any gas-borne suspended phase of a compound described herein which is capable of being inhaled into the bronchioles or nasal passages, and includes dry powder and aqueous aerosol, and pulmonary and nasal aerosols.
  • aerosol includes a gas-born suspension of droplets of a compound described herein, as may be produced in a metered dose inhaler or nebulizer, or in a mist sprayer.
  • Aerosol also includes a dry powder composition of a composition of the present technology suspended in air or other carrier gas, which may be delivered by insufflation from an inhaler device, for example.
  • a dry powder composition of a composition of the present technology suspended in air or other carrier gas which may be delivered by insufflation from an inhaler device, for example.
  • compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Any pharmaceutically acceptable dosage form may be employed in the methods of the present technology.
  • the composition can be formulated: (a) for administration selected from the group consisting of oral,-pulmonary, rectal, colonic, parenteral, intracisternal, intravaginal,
  • An exemplary dosage form is an orally administered dosage form, such as a tablet or capsule.
  • Such methods include the step of bringing into association Form 1 and/or Form 2 of squalamine phosphate with the carrier that constitutes one or more accessory ingredients.
  • the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
  • Formulations or compositions of the present technology may be packaged together with, or included in a kit with, instructions or a package insert.
  • instructions or package inserts may address recommended storage conditions, such as time, temperature and light, taking into account the shelf-life of Form 1 and/or Form 2 of squalamine phosphate.
  • Such instructions or package inserts may also address the particular advantages of Form 1 and/or Form 2 of squalamine phosphate, such as the ease of storage for formulations that may require use in the field, outside of controlled hospital, clinic or office conditions.
  • the composition disclosed herein can also be included in nutraceuticals.
  • the composition may be administered in natural products, including milk or milk product.
  • Form 1 and/or Form 2 of squalamine phosphate can also be provided in powder or tablet form, with or without other known additives, carriers, fillers and diluents.
  • Exemplary nutraceuticals are described in Scott Hegenhart, Food Product Design, December 1993.
  • composition disclosed herein will be formulated and dosed in a fashion consistent with good medical practice, taking into account the clinical condition of the individual patient (especially the side effects of treatment with the solid form(s) of squalamine phosphate alone), the site of delivery, the method of administration, the scheduling of administration, and other factors known to practitioners.
  • the "effective amount" for purposes herein is thus determined by such considerations.
  • the total pharmaceutically effective amount of Form 1 and/or Form 2 of squalamine phosphate administered parenterally per dose will be in the range of about 0.1 mg/kg/day to 20 mg/kg/day of patient body weight, although, as noted above, this will be subject to therapeutic discretion.
  • An intravenous bag solution may also be employed. The length of treatment needed to observe changes and the interval following treatment for responses to occur appears to vary depending on the desired effect.
  • “Pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
  • compositions disclosed herein can be formulated by combining Form 1 and/or Form 2 of squalamine phosphate with pharmaceutically acceptable carriers known in the art.
  • Such carriers enable Form 1 and/or Form 2 of squalamine phosphate to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated.
  • Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with Form 1 and/or Form 2 of squalamine phosphate, optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • coatings for solid oral formulations include sugar coating and film coating.
  • Film coatings may contain polymers such as, but not limited to, hydroxypropyl methylcellulose (HPMC), methyl hydroxyethyl cellulose, ethylcellulose, povidone, cellulose acetate phthalate, acrylate polymers (e.g., Eudragit L and Eudragit S), and HPMC phthalate.
  • Form 1 and/or Form 2 of squalamine phosphate is also suitably administered by sustained-release systems.
  • sustained-release compositions may be administered orally, rectally, parenterally, intracistemally, intravaginally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), bucally, or as an oral or nasal spray.
  • sustained-release compositions include suitable polymeric materials (such as, for example, semi-permeable polymer matrices in the form of shaped articles, e.g., films, or mirocapsules), suitable hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (such as, for example, a sparingly soluble salt).
  • suitable polymeric materials such as, for example, semi-permeable polymer matrices in the form of shaped articles, e.g., films, or mirocapsules
  • suitable hydrophobic materials for example as an emulsion in an acceptable oil
  • ion exchange resins for example as an emulsion in an acceptable oil
  • sparingly soluble derivatives such as, for example, a sparingly soluble salt
  • Sustained-release matrices include polylactides (U.S. Pat. No. 3,773,919, EP 58,481), copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman et al., Biopolymers, 22:547-556 (1983)), poly (2-hydroxyethyl methacrylate) (Langer et al., J Biomed. Mater. Res., 75: 167-277 (1981), and Langer, Chem. Tech, 72:98-105 (1982)), ethylene vinyl acetate (Langer et al., Id.) or poly-D-(-)-3-hydroxybutyric acid (EP 133,988).
  • polylactides U.S. Pat. No. 3,773,919, EP 58,481
  • copolymers of L-glutamic acid and gamma-ethyl-L-glutamate (Sidman et al
  • Sustained-release compositions may also include liposomally entrapped compounds described herein (see generally, Langer, Science, 249: 1527-1533 (1990); Treat et al, in
  • Liposomes comprising Form 1 and/or Form 2 of squalamine phosphate may be prepared by methods known per se: DE 3,218,121 ; Epstein et al., Proc. Natl. Acad. Sci. (USA), 52:3688-3692 (1985); Hwang et al, Proc. Natl. Acad. Sci. (USA), 77:40304034 (1980); EP 52,322; EP 36,676; EP 88,046; EP 143,949; EP 142,641 ; Japanese Pat.
  • the liposomes are of the small (about 200-800 Angstroms) unilamellar type in which the lipid content is greater than about 30 mol. percent cholesterol, the selected proportion being adjusted for the optimal therapeutic.
  • the present technology also provides a pharmaceutical pack or kit comprising one or more containers filled with the composition described herein.
  • Associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
  • Form 1 and/or Form 2 of squalamine phosphate may be employed in conjunction with other therapeutic compounds.
  • compositions according to the present technology may also comprise one or more binding agents, filling agents, lubricating agents, suspending agents, sweeteners, flavoring agents, preservatives, buffers, wetting agents, disintegrants, effervescent agents, and other excipients.
  • excipients are known in the art.
  • filling agents include lactose monohydrate, lactose anhydrous, and various starches;
  • binding agents include various celluloses and cross-linked
  • microcrystalline cellulose such as Avicel ® PH101 and Avicel ® PH102, microcrystalline cellulose, and silicified microcrystalline cellulose (ProSolv SMCCTM).
  • Suitable lubricants including agents that act on the flowability of the powder to be compressed, may include colloidal silicon dioxide, such as Aerosil ® 200, talc, stearic acid, magnesium stearate, calcium stearate, and silica gel.
  • colloidal silicon dioxide such as Aerosil ® 200, talc, stearic acid, magnesium stearate, calcium stearate, and silica gel.
  • sweeteners may include any natural or artificial sweetener, such as sucrose, xylitol, sodium saccharin, cyclamate, aspartame, and acesulfame.
  • flavoring agents are Magnasweet ® (trademark of MAFCO), bubble gum flavor, and fruit flavors, and the like.
  • preservatives include potassium sorbate, methylparaben, propylparaben, benzoic acid and its salts, other esters of parahydroxybenzoic acid such as butylparaben, alcohols such as ethyl or benzyl alcohol, phenolic compounds such as phenol, or quaternary compounds such as benzalkonium chloride.
  • Suitable diluents include pharmaceutically acceptable inert fillers, such as
  • microcrystalline cellulose lactose, dibasic calcium phosphate, saccharides, and/or mixtures of any of the foregoing.
  • diluents include microcrystalline cellulose, such as Avicel ® PH101 and Avicel ® PH102; lactose such as lactose monohydrate, lactose anhydrous, and Pharmatose ® DCL21 ; dibasic calcium phosphate such as Emcompress ® ; mannitol; starch;
  • Suitable disintegrants include lightly crosslinked polyvinyl pyrrolidone, corn starch, potato starch, maize starch, and modified starches, croscarmellose sodium, cross-povidone, sodium starch glycolate, and mixtures thereof.
  • effervescent agents include effervescent couples such as an organic acid and a carbonate or bicarbonate.
  • Suitable organic acids include, for example, citric, tartaric, malic, fumaric, adipic, succinic, and alginic acids and anhydrides and acid salts.
  • Suitable carbonates and bicarbonates include, for example, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, sodium glycine carbonate, L-lysine carbonate, and arginine carbonate.
  • sodium bicarbonate component of the effervescent couple may be present.
  • Clin Cancer Res 9(7): 2465-71 describes exemplary dosages for a 5-day continuous i.v. infusion every 3 weeks for treating advanced solid malignancies. Dose levels ranging from 6 to 700 mg/m(2)/day. Hepatotoxicity, characterized by brief, asymptomatic elevations in transaminases and hyperbilirubinemia, was the principal dose- limiting toxicity of squalamine. At 700 mg/m(2)/day, two of three patients developed grade 4 hyperbilirubinemia, which precluded further dose escalation. At 500 mg/m(2)/day, one of seven patients experienced dose-limiting grade 4 hyperbilirubinemia and grade 3 neurosensory changes, which resolved soon after treatment.
  • Squalamine pharmacokinetics were dose- proportional. At 500 mg/m(2)/day, the mean (percentage coefficient of variation) clearance, half-life, and volume of distribution of squalamine were 2.67 liters/h/m(2) (85%), 9.46 h (81 %), and 36.84 liters/m(2) (124%), respectively, and steady-state concentrations [20.08 micro g/ml (13%)] were well above those that inhibit angiogenesis in preclinical models. The study concluded that at a dose of 500 mg/m(2)/day, squalamine is well tolerated.
  • squalamine Patients with chemotherapy-naive non- small cell lung cancer were treated with escalating doses of squalamine in combination with standard doses of paclitaxel and carboplatin.
  • Paclitaxel and carboplatin were administered on day 1, followed by squalamine as a continuous infusion on days 1 -5, every 21 days.
  • the starting dose of squalamine was 100 mg/m(2)/day and escalated to 400 mg/m(2)/day; two of three patients at 400 mg/m(2)/day had dose-limiting toxicity that included grade 3/4 arthralgia, myalgia, and neutropenia.
  • 300 mg/m(2)/day was selected as the Phase II dose of squalamine in this combination regimen.
  • the combination of squalamine given continuously daily for 5 days, with paclitaxel and carboplatin given on day 1 was well tolerated.
  • Form 1 and/or Form 2 of squalamine phosphate may be administered alone or in combination with one or more adjuvants.
  • an adjuvant is a substance that indirectly enhances the therapeutic activity of the squalamine phosphate by stimulating the antiviral arm of the innate and/or the adaptive immune system.
  • Adjuvants that may be administered with Form 1 and/or Form 2 of squalamine phosphate include, but are not limited to, cytokines and/or interleukins (such as IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL-9, 1L10, IL-1 1 , IL12, IL13, IL-14, IL15, IIL16, IL-17, IL-18, IL- 19, IL-20, IL-21 , anti-CD40, CD40L, IFN-gamma, TNF-alpha, IL-Ialpha, IL-lbeta), Lipid A, including monophosphoryl lipid A, bacterial products, endotoxins, cholesterol, fatty acids, aliphatic amines, paraffinic and vegetable oils, threonyl derivative, and muramyl dipeptide, alum, alum plus deoxycholate (ImmunoAg), MTP-PE (Biocine Corp.), QS21 (Genen
  • Form 1 and/or Form 2 of squalamine phosphate is administered in combination with alum. In another specific embodiment, Form 1 and/or Form 2 of squalamine phosphate is administered in combination with QS-21.
  • Further adjuvants that may be administered with Form 1 and/or Form 2 of squalamine phosphate include, but are not limited to, Monophosphoryl lipid immunomodulator, AdjuVax 100a, QS-21 , QS-18, CRL1005, Aluminum salts, MF-59, and Virosomal adjuvant technology.
  • Vaccines that may be administered with Form 1 and/or Form 2 of squalamine phosphate include any antigen capable of eliciting an immune response.
  • the vaccine may be comprised of either live or inactivated virus.
  • Exemplary vaccines include, but are not limited to, vaccines directed toward protection against MMR (measles, mumps, rubella), polio, varicella, tetanus/diptheria, hepatitis A, hepatitis B, Haemophilus influenzae B, whooping cough, pneumonia, influenza, Lyme's Disease, rotavirus, cholera, yellow fever, Japanese encephalitis, poliomyelitis, rabies, typhoid fever, pertussis, PA-toxin (e.g., anthrax), Human
  • HIV-1 and HIV-2 Immunodeficiency Virus
  • Avian Flu antigen e.g., H5N1 ; avian influenza virus A/FPV/Rostock/34 (H7N1) (FPV)
  • SARS Severe Acute Respiratory Syndrome
  • Useful antigens include but are not limited to viral, prion, bacterial, parasitic, mycotic, etc. antigens.
  • Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently; or sequentially.
  • “combination administration” includes compounds which are attached to Form 1 and/or Form 2 of squalamine phosphate This also includes presentations in which the combined agents are administered together as a therapeutic mixture, and also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same individual. Administration “in combination” further includes the separate administration of one of the compounds or agents given first, followed by the second.
  • the squalamine phosphate compositions of the present technology may be administered alone or in combination with other therapeutic agents.
  • the squalamine phosphate compositions of the present technology are useful in treating and/or preventing: (1) viral infections, (2) antimicrobial infections, including but not limited to Gram-negative and Gram-positive bacterial infections, fungal infections, and protozoan infections; (3) disease states known to be associated with pathological neovascularization, such as cancer, due to
  • squalamine' s anti-angiogenic properties vascular disorders of the eye, including macular degeneration, such as age-related macular degeneration, retinopathy of prematurity, corneal neovascularization, diabetic retinopathy; (5) weight loss or weight management, dose-dependent weight loss; (6) diseases, including viral infections, where sodium-hydrogen exchanger ("NHE- 3") plays a critical role, and where its inhibition (by squalamine) could be effected; and (7) treatment of fibrodysplasia ossificans progressiva, a rare disease where connective tissue will ossify when damaged.
  • any active agent known to be useful in treating these conditions can be used in conjunction with the squalamine phosphate compositions of the present technology.
  • compositions of the present technology can be co-administered or combined with an antibiotic.
  • the squalamine phosphate compositions of the present technology can be co-administered or combined with an antiviral agent, etc.
  • the squalamine phosphate compositions described herein may be administered in combination compounds including but not limited to, chemotherapeutic agents, antibiotics, steroidal and non-steroidal anti-inflammatories, conventional immunotherapeutic agents, antiviral agents, and/or therapeutic treatments described below. Combinations may be administered either concomitantly, e.g., as an admixture, separately but simultaneously or concurrently; or sequentially.
  • Administration "in combination” further includes the separate administration of one of the compounds or agents given first, followed by the second.
  • squalamine phosphate compositions of the present technology can be combined with conventional antiviral therapies for treating and preventing viral infections.
  • the squalamine phosphate compositions of the present technology can be combined with any known antiviral agent.
  • Conventional antiviral treatments include, but are not limited to (1) Amantadine and rimantadine, which combat influenza and act on penetration/uncoating; (2) Pleconaril, which works against rhinoviruses, which cause the common cold; (3) nucleotide or nucleoside analogues, such as acyclovir, zidovudine (AZT), lamivudine; (4) drugs based on "antisense” molecules, such as fomivirsen; (5) ribozyme antivirals; (6) protease inhibitors; (7) assembly inhibitors, such as Rifampicin; (8) release phase inhibitors, such as zanamivir (Relenza) and oseltamivir (Tamiflu); (9) drugs which stimulate the immune system, such as interferons, which inhibit viral synthesis in infected cells (e.g., interferon alpha), and synthetic antibodies (A monoclonal drug is now being sold to help fight respiratory syncytial virus
  • antiviral drugs include, but are not limited to, Abacavir, Aciclovir, Acyclovir, Adefovir, Amantadine, Amprenavir, Arbidol, Atazanavir, Atripla, Boceprevir, Cidofovir, Combivir, Darunavir, Delavirdine, Didanosine, Docosanol, Edoxudine, Eiavirenz, Emtricitabine, Enfuvirtide, Entecavir, Entry inhibitors, Famciclovir, Fixed dose combination (antiretro viral), Fomivirsen, Fosamprenavir, Foscarnet, Fosfonet, Fusion inhibitor, Ganciclovir, Ibacitabine, Imunovir, Idoxuridine, Imiquimod, Indinavir, Inosine, Integrase inhibitor, Interferon type III, Interferon type II, Interferon type I, Interferon, Lamivudine, Lopinavir, Loviride,
  • Podophyllotoxin Protease inhibitor (pharmacology), Raltegravir, Reverse transcriptase inhibitor, Ribavirin, Rimantadine, Ritonavir, Saquinavir, Stavudine, Synergistic enhancer (antiretroviral), Tenofovir, Tenofovir disoproxil, Tipranavir, Trifluridine, Trizivir, Tromantadine, Truvada, Valaciclovir (Valtrex), Valganciclovir, Vicriviroc, Vidarabine, Viramidine, Zalcitabine,
  • Form 1 and/or Form 2 of squalamine phosphate is
  • NRTIs transcriptase inhibitors
  • NRTls non-nucleoside reverse transcriptase inhibitors
  • Pis protease inhibitors
  • NRTIs that may be administered in combination with Form 1 and/or Form 2 of squalamine phosphate, include, but are not limited to, RETROVIRTM (zidovudine/AZT), VIDEXTM (didanosinelddi), HIVIDTM (zalcitabine/ddC), ZERITTM
  • NNRTls that may be administered in combination with squalamine composition, include, but are not limited to, VIRAMUNETM (nevirapine), RESCRIPTORTM (delavirdine), and SUSTIVATM (efavirenz).
  • Protease inhibitors that may be administered in combination with Form 1 and/or Form 2 of squalamine phosphate include, but are not limited to, CRIXIVANTM (indinavir), NORVIRTM (ritonavir), INVIRASETM (saquinavir), and VIRACEPTTM (nelfinavir).
  • antiretroviral agents nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, and/or protease inhibitors may be used in any combination with Form 1 and/or Form 2 of squalamine phosphate to treat AIDS and/or to prevent or treat HIV infection.
  • Additional NRTIs include LODENOSINETM (F-ddA; an acid-stable adenosine NRTI; Triangle/Abbott; COVIRACILTM (emtricitabine/FTC; structurally related to lamivudine (3TC) but with 3-to 10-fold greater activity in vitro; Triangle/Abbott); dOTC (BCH- 10652, also structurally related to lamivudine but retains activity against a substantial proportion of lanivudine -resistant isolates; Biochem Pharma); Adefovir (refused approval for anti-HIV therapy by FDA; Gilead Sciences); PREVEON ® (Adefovir Dipivoxil, the active prodrug of adefovir; its active form is PMEA-pp); TENOFOVIRTM (bis-POC PMPA, a PMPA prodrug; Gilead);
  • DAPD/DXG active metabolite of DAPD; Triangle/Abbott
  • D-D4FC related to 3TC, with activity against AZT/3TC-resistant virus
  • GW420867 Gaxo Wellcome
  • Additional NNRTls include COACTINONTM (Emivirine/M C442, potent NNRTI of the HEPT class; Triangle/Abbott); CAPRAVIRINETM (AG-1549/S-1 153, a next generation NNRTI with activity against viruses containing the K103N mutation; Agouron); PNU-142721 (has 20- to 50-fold greater activity than its predecessor delavirdine and is active against K103N mutants; Pharmacia & Upjohn); DPC-961 and DPC-963 (second-generation derivatives of efavirenz, designed to be active against viruses with the K103N mutation; DuPont);
  • GW420867X has 25-fold greater activity than HBY097 and is active against K103N mutants; Glaxo Wellcome); CALANOLIDE A (naturally occurring agent from the latex tree; active against viruses containing either or both the Y181C and K103N mutations); and Propolis (WO 99/49830).
  • Additional protease inhibitors include LOPINAVIRTM (ABT378/r; Abbott
  • BMS-232632 an azapeptide; Bristol-Myres Squibb
  • TIPRANAVIRTM PNU- 140690, a non-peptic dihydropyrone; Pharmacia & Upjohn
  • PD-178390 a nonpeptidic dihydropyrone; Parke-Davis
  • BMS 232632 an azapeptide; Bristol-Myers Squibb
  • L-756,423 an indinavir analog; Merck
  • DMP450 a cyclic urea compound; Avid & DuPont
  • AG- 1776 a peptidomimetic with in vitro activity against protease inhibitor-resistant viruses; Agouron
  • VX- 175/GW433908 phosphate prodrug of amprenavir; Vertex & Glaxo Welcome
  • CGP61755 Ciba
  • AGENERASETM amprenavir; Glaxo Wellcome Inc.
  • Additional antiretroviral agents include fusion inhibitors/gp41 binders.
  • Fusion inhibitors/gp41 binders include T-20 (a peptide from residues 643-678 of the HIV gp41 transmembrane protein ectodomain which binds to gp41 in its resting state and prevents transformation to the fusogenic state; Trimeris) and T-1249 (a second-generation fusion inhibitor; Trimeris).
  • Additional antiretroviral agents include fusion inhibitors/chemokine receptor antagonists.
  • Fusion inhibitors/chemokine receptor antagonists include CXCR4 antagonists such as AMD 3100 (a bicyclam), SDF-1 and its analogs, and ALX404C (a cationic peptide), T22 (an 18 amino acid peptide; Trimeris) and the T22 analogs T134 and T140; CCR5 antagonists such as RANTES (9-68), AOP-RANTES, NNY-RANTES, and TAK-779; and CCR5/CXCR4 antagonists such as NSC 651016 (a distamycin analog). Also included are CCR2B, CCR3, and CCR6 antagonists. Chemokine receptor agonists such as RANTES, SDF-1, MEP-1 alpha, ⁇ - lbeta, etc., may also inhibit fusion.
  • Additional antiretroviral agents include integrase inhibitors.
  • Integrase inhibitors include dicaffeoylquinic (DFQA) acids; L-chicoric acid (a dicaffeoyl tartaric (DCTA) acid); quinalizarin (QLC) and related anthraquinones; ZINTEVIRTM (AR 177, an oligonucleotide that probably acts at cell surface rather than being a true integrase inhibitor; Arondex); and naphthols such as those disclosed in WO 98/50347.
  • DFQA dicaffeoylquinic
  • DCTA dicaffeoyl tartaric
  • QLC quinalizarin
  • ZINTEVIRTM AR 177, an oligonucleotide that probably acts at cell surface rather than being a true integrase inhibitor
  • Arondex naphthols such as those disclosed in WO 98/50347.
  • Additional antiretroviral agents include hydroxyurea-like compounds such as BCX-34 (a purine nucleoside phosphorylase inhibitor; Biocryst); ribonucleotide reductase inhibitors such as DIDOXTM (Molecules for Health); inosine monophosphate dehydrogenase (IMPDH) inhibitors such as VX-497 (Vertex); and mycopholic acids such as CellCept (mycophenolate mofetil; Roche).
  • BCX-34 purine nucleoside phosphorylase inhibitor
  • ribonucleotide reductase inhibitors such as DIDOXTM (Molecules for Health)
  • IMPDH inosine monophosphate dehydrogenase
  • VX-497 Verytex
  • mycopholic acids such as CellCept (mycophenolate mofetil; Roche).
  • Additional antiretroviral agents include inhibitors of viral integrase, inhibitors of viral genome nuclear translocation such as arylene bis(methylketone) compounds; inhibitors of HIV entry such as AOP-RANTES, NNY-RANTES, RANTES-IgG fusion protein, soluble complexes of RANTES and glycosaminoglycans (GAG), and AMD-3100; nucleocapsid zinc finger inhibitors such as dithiane compounds; targets of HIV Tat and Rev; and pharmacoenhancers such as ABT-378.
  • inhibitors of viral integrase inhibitors of viral genome nuclear translocation such as arylene bis(methylketone) compounds
  • inhibitors of HIV entry such as AOP-RANTES, NNY-RANTES, RANTES-IgG fusion protein, soluble complexes of RANTES and glycosaminoglycans (GAG), and AMD-3100
  • nucleocapsid zinc finger inhibitors such as dithiane compounds
  • cytokines and lymphokines such as MIP-lalpha, MIP-lbeta, SDF-lalpha, IL-2, PROLEUKINTM (aldesleukin/L2-7001 ; Chiron), IL4, IL-10, IL-12, and IL-13; interferons such as IFN-alpha2a, IFN-alpha2b, or IFN- beta; antagonists of TNFs, NFkappaB, GM-CSF, M-CSF, and IL-10; agents that modulate immune activation such as cyclosporin and prednisone; vaccines such as RemuneTM (HIV Immunogen), APL 400-003 (Apollon), recombinant g l20 and fragments, bivalent (B/E) recombinant envelope glycoprotein, rgpl20CM235, MN rgpl20, SF-2 rgpl20, g l20/
  • antibodies such as the anti-CXCR4 antibody 12G5, the anti-CCR5 antibodies 2D7, 5C7, PA8, PA9, PA10, PA1 1 , PA12, and PA14, the anti-CD4 antibodies Q4120 and RPA-T4, the anti-CCR3 antibody 7B1 1 , the anti-gpl20 antibodies 17b, 48d, 447-52D, 257-D, 268-D and 50.1 , anti-Tat antibodies, anti-TNF-alpha antibodies, and monoclonal antibody 33 A; aryl hydrocarbon (AH) receptor agonists and antagonists such as TCDD, 3,3',4,4',5- pentachlorobiphenyl, 3,3',4,4'-tetrachlorobiphenyl, and alpha-naphthoflavone (WO 98/30213); and antioxidants such as garnma-L-glutamyl-L-cysteine ethyl ester (
  • the squalamine phosphate composition described herein is administered alone or in combination with an anti-inflammatory agent.
  • Anti-inflammatory agents that may be administered with the squalamine phosphate compositions described herein include, but are not limited to, corticosteroids (e.g., betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone), nonsteroidal anti-inflammatory drugs (e.g., diclofenac, diflunisal, etodolac, fenoprofen, floctafenine, flurbiprofen, ibuprofen, indomethacin, ketoprofen, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, pir
  • aminoarylcarboxylic acid derivatives arylacetic acid derivatives, arylbutyric acid derivatives, arylcarboxylic acids, arylpropionic acid derivatives, pyrazoles, pyrazolones, salicylic acid derivatives, thiazinecarboxamides, e-acetamidocaproic acid, S-adenosylmethionine, 3-amino4- hydroxybutyric acid, amixetrine, bendazac, benzydamine, bucolome, difenpiramide, ditazol, emorfazone, guaiazulene, nabumetone, nimesulide, orgotein, oxaceprol, paranyline, perisoxal, pifoxime, proquazone, proxazole, and tenidap.
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more antibiotics.
  • antibiotics include, but are not limited to, aminoglycosides, Ansamycins, Carbacephems,
  • Carbapenems Cephalosporins, Glycopeptides, Macrolides, Monobactams, Penicillins, Polypeptides, Polymyxin, Quinolones, Sulfonamides, Tetracyclines, and others (e.g., Arsphenamine, Chloramphenicol, Clindamycin, Lincomycin, Ethambutol, Fosfomycin, Fusidic acid, Furazolidone, Isoniazid, Linezolid, Metronidazole, Mupirocin, Nitrofurantoin,
  • antibiotics include, but are not limited to, Amikacin, Gentamicin, Kanamycin, Neomycin, Netilmicin, Streptomycin, Tobramycin, Paromomycin, Geldanamycin, Herbimycin, Loracarbef, Ertapenem, Doripenem, Imipenem/Cilastatin,
  • Temafloxacin Mafenide, Sulfonamidochrysoidine (archaic), Sulfacetamide, Sulfadiazine, Sulfamethizole, Sulfanilimide (archaic), Sulfasalazine, Sulfisoxazole, Trimethoprim, rimethoprim-SuIfamethoxazole (Co-trimoxazole) (TMP-SMX), Demeclocycline, Doxycycline, Minocycline, Oxytetracycline, and Tetracycline.
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more antifungal, antiyeast or antimold agents.
  • Exemplary active agents include, but are not limited to, (1) azoles (imidazoles), (2) antimetabolites, (3) allylamines, (4) morpholine, (5) glucan Synthesis Inhibitors (chemical family: echinocandins), (6) polyenes, (7) benoxaborales, (8) other antifungal/onychomycosis agents, and (9) new classes of antifungal/onychomycosis agents.
  • Examples of azoles include, but are not limited to, Bifonazole, Clotrimazole,
  • An example of an antimetabolite includes, but is not limited to, Flucytosine.
  • Examples of allylamines include, but are not limited to,
  • glucan Synthesis Inhibitors include, but are not limited to, Caspofungin, Micafungin, and
  • Anidulafungin examples include, but are not limited to, Amphotericin B, Nystatin, and pimaricin.
  • An example of a benoxaborale is AN2690.
  • Other examples of antifungal agents include, but are not limited to, griseofulvin and ciclopirox.
  • examples of new classes of antifungal/onychomycosis agents include, but are not limited to, sodarin derivatives and nikkomycins.
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more anticancer agents.
  • the anticancer agent may be a nitrosourea (such as BCNU), cyclophosphamide, adriamycin, 5- fluorouracil, paclitaxel and its derivatives, cisplatin or other platinum containing cancer treating agents.
  • chemotherapeutic agent that can be used in this present technology.
  • chemotherapeutic agents that can be used with the squalamine phosphate composition of the present technology include, for example, methotrexate, thiotepa, mitoxantrone, vincristine, vinblastine, etoposide, ifosfamide, bleomycin, procarbazine, chlorambucil, fludarabine, mitomycin C, vinorelbine, and gemcitabine.
  • chemotherapeutic drugs can be divided in to alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antitumour agents. All of these drugs affect cell division or DNA synthesis and function in some way. Some newer agents do not directly interfere with DNA. These include monoclonal antibodies and the new tyrosine kinase inhibitors e.g. imatinib mesylate (Gleevec or Glivec), which directly targets a molecular abnormality in certain types of cancer (chronic myelogenous leukemia, gastrointestinal stromal tumors). In addition, some drugs that modulate tumor cell behaviour without directly attacking those cells may be used. Hormone treatments fall into this category.
  • Alkylating agents Alkylating antineoplastic agents are so named because of their ability to add alkyl groups to many electronegative groups under conditions present in cells. Cisplatin and carboplatin, as well as oxaliplatin, are alkylating agents. Other agents are mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide.
  • Anti-metabolites Plant alkaloids and terpenoids are derived from plants and block cell division by preventing microtubule function.
  • the main examples are vinca alkaloids and taxanes.
  • Vinca alkaloids include Vincristine, Vinblastine, Vinorelbine, and Vindesine.
  • Podophyllotoxin is used to produce two other cytostatic drugs, etoposide and teniposide.
  • the prototype taxane is the natural product paclitaxel.
  • Docetaxel is a semi-synthetic analogue of paclitaxel.
  • Topoisomerase inhibitors include camptothecins (irinotecan and topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, dactinomycin, doxorubicin, epirubicin, bleomycin and others.
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more appetite suppressants or weight loss agents.
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more agents targeting a neurological condition.
  • the methods of the invention are useful in treating and/or preventing the conditions described herein, including but not limited to neurological disorders, such as Parkinson's disease, autism, multiple system atrophy, depression, Alzheimer's disease, Huntington's Disease, schizophrenia, multiple sclerosis, and degenerative processes associated with aging, autonomic system instability, including delays in sleep onset, fragmentation of sleep, reduced REM sleep, reduced total sleep time, REM-behavior disorder, sleep breathing disorder including snoring and sleep apnea, hallucinations, narcolepsy, and day-time sleepiness.
  • neurological disorders such as Parkinson's disease, autism, multiple system atrophy, depression, Alzheimer's disease, Huntington's Disease, schizophrenia, multiple sclerosis, and degenerative processes associated with aging, autonomic system instability, including delays in sleep onset, fragmentation of sleep, reduced REM sleep, reduced total sleep time, REM-
  • the squalamine phosphate composition described herein is administered alone or in combination with one or more agents used to treat gastronintestinal disorders such as constipation, Crohn's disease or IBS.
  • any active agent known to be useful in treating these conditions can be used in the disclosed, and either combined with the aminosterol compositions used in the methods, or administered separately or sequentially.
  • the present technology also provides methods of treatment and/or prevention of diseases or disorders (such as, for example, any one or more of the diseases or disorders disclosed herein) by administration to a subject of an effective amount of Form 1 and/or Form 2 of squalamine phosphate in one or more pharmaceutically acceptable carriers.
  • the compositions of the present technology can be administered using any pharmaceutically acceptable method, including but not limited to oral, pulmonary, nasal, and nebularization administration.
  • the squalamine phosphate compositions of the present technology can be used to treat any indication known to be amenable to treatment with squalamine.
  • the squalamine phosphate compositions of the present technology can be used to treat, for example, (1) viral infections, (2) antimicrobial infections, including but not limited to treating and/or preventing Gram-negative and Gram-positive bacterial infections, fungal infections, and protozoan infections; (3) disease states known to be associated with pathological neovascularization, such as cancer, due to squalamine's anti-angiogenic properties; (4) vascular disorders of the eye, including macular degeneration, such as age-related macular degeneration, retinopathy of prematurity, corneal neovascularization, diabetic retinopathy; (5) weight loss or weight management, dose-dependent weight loss; (6) diseases, including viral infections, where sodium-hydrogen exchanger ("NHE- 3") plays a critical role, and where its inhibition (by squalamine) could be effected; (7) treatment of fibr
  • Alzheimer's disease Huntington's Disease, acute traumatic injury to the central nervous system, including the spinal cord, stroke, acute head and/or spine injury, degenerative processes associated with aging, including memory loss ("dementia of aging"), cerebral palsy, epilepsy, peripheral sensory neuropathy, and multiple sclerosis;(12) autism; (13) sleep disorders; (14) schizophrenia; (15) depression; and (16) autonomic system lability.
  • provided herein are methods for treating a subject in need having a condition susceptible to treatment with an aminosterol, comprising
  • a composition comprising or consisting essentially of a squalamine phosphate solid form of Form 1 and/or Form 2, and one or more pharmaceutically acceptable carriers and/or excipients.
  • the subject in need has a condition selected from the group consisting of viral infections, antimicrobial infections, Gram-negative and Gram-positive bacterial infections, Mycobacteria infections, fungal infections, protozoan infections, disease states known to be associated with pathological neovascularization, such as cancer, vascular disorders of the eye, including macular
  • the viral infection is caused by a virus selected from the group consisting of Yellow Fever, Cytomegalovirus, Eastern Equine Encephalitis virus, Hepatitis B virus, Hepatitis Delta virus, Dengue virus, and Human Immunodeficiency virus.
  • the condition to be treated is a viral infection caused by a virus selected from the group consisting of "African Swine Fever Viruses," Arbovirus, Adenoviridae,
  • Arenaviridae Arterivirus, Astroviridae, Baculoviridae, Bimaviridae, Birnaviridae, Bunyaviridae, Caliciviridae, Caulimoviridae, Circoviridae, Coronaviridae, Cystoviridae, Dengue, EBV, HIV, Deltaviridae, Filviridae, Filoviridae, Flaviviridae, Hepadnaviridae (Hepatitis), Herpesviridae (such as, Cytomegalovirus, Herpes Simplex, Herpes Zoster), Iridoviridae, Mononegavirus (e.g., Paramyxoviridae, Morbillivirus, Rhabdoviridae), Myoviridae, Orthomyxoviridae (e.g., Influenza A, Influenza B, and parainfluenza), Papiloma virus, Papovaviridae, Paramyxoviridae, Prions,
  • the condition to be treated is selected from the group consisting of AIDS, viral meningitis, Dengue, EBV, hepatitis, a chronic disease suspected to be of viral origin, multiple sclerosis, Type I diabetes, Type II diabetes, atherosclerosis, cardiomyopathies, Kawaski disease, aplastic anemia, and any combination thereof.
  • the method further comprises
  • an additional active agent wherein the additional active agent is administered via a method selected from the group consisting of (a) concomitantly; (b) as an admixture; (c) separately and simultaneously or concurrently; and (d) separately and sequentially.
  • the subject is human.
  • kits for treating and/or preventing viral infections comprising administering a therapeutically effective amount of a squalamine phosphate composition disclosed herein to a subject in need.
  • a "subject in need” is a human or animal at risk of a viral infection, or which has contracted a viral infection.
  • this method encompasses using a squalamine phosphate composition described herein in combination with conventional antiviral treatments to treat viral infections.
  • the viral infection to be treated or prevented can be caused by any virus, including but not limited to, "African Swine Fever Viruses,” Arbovirus, Adenoviridae, Arenaviridae,
  • Paramyxoviridae Morbillivirus, Rhabdoviridae), Myoviridae, Orthomyxoviridae (e.g., Influenza A, Influenza B, and parainfluenza), Papiloma virus, Papovaviridae, Paramyxoviridae, Prions, Parvoviridae, Phycodnaviridae, Picomaviridae (e.g., Rhinovirus, Poliovirus), Poxviridae (such as Smallpox or Vaccinia), Potyviridae, Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-1, HTLV- II, Lentivirus), Rhabdoviridae, Tectiviridae, Togaviridae (e.g., Rubivirus), or any combination thereof.
  • Reoviridae e.g., Rotavirus
  • Retroviridae HTLV-1, HTLV- II, Lenti
  • the viral infection is caused by a virus selected from the group consisting of herpes, pox, papilloma, corona, influenza, hepatitis, sendai, Sindbis, vaccinia viruses, west nile, hanta, or viruses which cause the common cold.
  • the condition to be treated is selected from the group consisting of AIDS, viral meningitis, Dengue, EBV, hepatitis, and any combination thereof.
  • the condition to be treated is a chronic disease suspected to be of viral origin.
  • the condition to be treated can be multiple sclerosis, Type I diabetes, Type II diabetes, atherosclerosis, cardiomyopathies, Kawaski disease, aplastic anemia, etc.
  • combination methods of treating or preventing a viral infection comprise:
  • squalamine composition and conventional antiviral drug can be administered sequentially or simultaneously. If Form 1 and/or Form 2 of squalamine phosphate or a conventional antiviral drug are administered sequentially, either squalamine or the conventional antiviral drug can be administered first.
  • viruses are not expected to develop resistance to squalamine. This is because unlike conventional antiviral therapies, squalamine does not act upon a single mechanism by which a virus infects a cell.
  • squalamine changes the cell structure for a period of time during which the virus cannot infect the cell.
  • certain anti-HIV drugs target the CD4 receptor and other antiviral drugs target inhibition of replication.
  • Viral variants can circumvent each of these targeted antiviral therapies.
  • squalamine does not demonstrate an altered IC50 or IC90 (drug concentration required to inhibit viral growth by 50% or 90% respectively) over time.
  • squalamine demonstrates an IC50 or IC90 which does not increase by more than 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 1 1%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, or 30% over time.
  • the time period over which the change in IC50 or IC90 (or lack thereof) is measured is 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 1 .5 months, 2 months, 2.5 months, 3 months, 3.5 months, 4 months, 4.5 months, 5 months, 5.5 months, 6 months, 6.5 months, 7 months, 7.5 months, 8 months, 8.5 months, 9 months, 9.5 months, 10 months, 10.5 months, 1 1 months, 1 1.5 months, 12 months, 1 year, 1.5 years, 2 years, 2.5 years, 3 years, 3.5 years, 4 years, 4.5 years, or 5 years.
  • Toxicity Conventional antiviral agents are generally designed to target viral specific enzymes, such as RNA and DNA polymerases, proteases, or glycosidases; and as a consequence the drug inhibits the activity of the viral enzyme to a far greater extent than it does to analogous human enzymes, required for normal cellular functioning. In many instances toxicity develops as a consequence of the residual activity of the agent towards the analogous enzymes of the host.
  • viral specific enzymes such as RNA and DNA polymerases, proteases, or glycosidases
  • the present technology is directed to methods of treating and/or preventing microbial infections, and in particular pathogenic microorganisms, comprising administering a therapeutically effective amount of a squalamine phosphate composition described herein to a subject in need.
  • this method encompasses using a squalamine phosphate composition described herein in combination with conventional antimicrobial treatments to treat and/or prevent infections.
  • a "subject in need" is a human or animal at risk of a microbial infection, or which has contracted a microbial infection.
  • microorganism refers to microscopic organisms and taxonomically related macroscopic organisms within the categories of algae, bacteria, fungi (including lichens), protozoa, viruses, and subviral agents.
  • microorganism refers to microscopic organisms and taxonomically related macroscopic organisms within the categories of algae, bacteria, fungi (including lichens), protozoa, viruses, and subviral agents.
  • microorganism refers to microscopic organisms and taxonomically related macroscopic organisms within the categories of algae, bacteria, fungi (including lichens), protozoa, viruses, and subviral agents.
  • pathogenic encompasses both those organisms that are in and of themselves pathogenic to another organism (e.g., animals, including humans, and plants) and those organisms that produce agents that are pathogenic to another organism, while the organism itself is not directly pathogenic or infective to the other organism.
  • pathogen refers to an organism, including microorganisms, that causes disease in another organism (e.g., animals and plants) by directly infecting the other organism, or by producing agents that causes disease in another organism (e.g., bacteria that produce pathogenic toxins and the like).
  • the bacterial infection to be treated and/or prevented can be due to a gram negative bacteria, gram positive bacteria, Mycobacteria, bacterial spore, or any combination thereof.
  • Pathogenic bacteria are a major cause of human death and disease and cause infections such as tetanus, typhoid fever, diphtheria, syphilis, cholera, foodborne illness, leprosy and tuberculosis.
  • gram positive bacteria include, but are not limited to genera such as
  • Staphylococcus Streptococcus, Enterococcus, (which are cocci) and Bacillus, Corynebacterium, Nocardia, Clostridium, Actinobacteria, and Listeria.
  • gram negative bacteria include, but are not limited to, Escherichia coli, Salmonella, Shigella, Enter obacteriaceae, Neisseria, Pseudomonas, Moraxella, Helicobacter, Stenotrophomonas, Bdellovibrio, acetic acid bacteria, Legionella and alpha-proteobacteria as Wolbachia and many others.
  • Gram-negative bacteria include the cyanobacteria, spirochaetes, green sulfur and green non-sulfur bacteria.
  • Medically relevant Gram-negative bacilli include a multitude of species. Some of them primarily cause respiratory problems ⁇ Hemophilus influenzae, Klebsiella pneumoniae, Legionella pneumophila, Pseudomonas aeruginosa), primarily urinary problems ⁇ Escherichia coli, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens), and primarily gastrointestinal problems ⁇ Helicobacter pylori, Salmonella enteritidis, Salmonella typhi).
  • Gram- negative bacteria associated with nosocomial infections include Acinetobacter baumannii, which cause bacteremia, secondary meningitis, and ventilator-associated pneumonia in intensive care units of hospital establishments.
  • Relevant Mycobacteria include M. tuberculosis complex (MTBC), M. tuberculosis, M. bovis, M. ajricanum, and M. microti; M. leprae, M. avium complex, M. avium paraluberculosis, M. avium sylvaticum, or any of the Mycobacterial species demonstrated to cause disease in man and/or animals.
  • the fungal, yeast and/or mold infection to be treated, prevented, and/or cured may be a tinea infection, dermatophytoses, or a dermatophytoma.
  • fungal microorganisms include, but are not limited to, Trichophyton spp. , Epidermophylon spp., Fusarium spp.,
  • Molds include, but are not limited to infections caused by the fungi Acremonium spp., Aspergillus spp. (e.g., A. sydowii, A. terreus, A. niger), Fusarium spp. (e.g., F. oxysporum, F. solani, F. semitectum), Scopulariopsis spp. (e.g., Scopulariopsis brevicaulis), Scedosporuim spp., Alternaria spp., Paecilomyces lilacinus, Epiccocum nigrum, Phoma spp. Chaetomium spp., Curvularia spp., Onychocola canadensis, and Scytalidium spp., (e.g., S. dimidiatum).
  • Aspergillus spp. e.g., A. sydowii, A. terreus, A.
  • Yeast as defined herein, include, but are not limited to, Candida species causing yeast infections.
  • the present technology is directed to methods of treating malignant and cancerous tumors, comprising administering a therapeutically effective amount of a squalamine phosphate composition described herein to a subject in need.
  • a "subject in need” is a human or animal having a malignant and cancerous tumor.
  • this method encompasses using a squalamine phosphate composition described herein in combination with conventional cancer treatments to treat tumors.
  • Examples of tumors that can be treated with the compositions of the present technology include, but are not limited to, breast, brain, lung (e.g., non-small cell lung cancer), and CNS.
  • An example of a solid brain tumor that can be treated with a composition according to the present technology is a malignant glioma.
  • Other examples of cancers that can be treated with compositions according to the present technology include, but are not limited to, prostate cancer.
  • the invention also encompasses use of the pharmaceutical compositions of the invention in methods of treatment of neurological disorders, such as Parkinson's disease, autism, multiple system atrophy, depression, Alzheimer's disease, Huntington's Disease, schizophrenia, multiple sclerosis, and degenerative processes associated with aging, autonomic system instability, including delays in sleep onset, fragmentation of sleep, reduced REM sleep, reduced total sleep time, REM-behavior disorder, sleep breathing disorder including snoring and sleep apnea, hallucinations, narcolepsy, and day-time sleepiness.
  • neurological disorders such as Parkinson's disease, autism, multiple system atrophy, depression, Alzheimer's disease, Huntington's Disease, schizophrenia, multiple sclerosis, and degenerative processes associated with aging, autonomic system instability, including delays in sleep onset, fragmentation of sleep, reduced REM sleep, reduced total sleep time, REM-behavior disorder, sleep breathing disorder including snoring and sleep apnea, hallucinations, narcolepsy,
  • the invention also encompasses use of the pharmaceutical compositions of the invention in methods of treatment of gastronintestinal disorders such as constipation, Crohn's disease or IBS.
  • any active agent known to be useful in treating these conditions can be used in the disclosed, and either combined with the aminosterol compositions used in the methods, or administered separately or sequentially.
  • a process for preparing the squalamine phosphate solid form of Form 1 comprising: (a) combining a solution of squalamine lactate with an aqueous solution of sodium phosphate dibasic and sodium phosphate monobasic to form a combined mixture; (b) heating the combined mixture before allowing the combined mixture to cool; and (c) isolating the combined mixture to isolate the squalamine phosphate solid form.
  • squalamine lactate refers to any lactate salt of squalamine. In some embodiments, squalamine lactate refers to squalamine monolactate. In some embodiments, squalamine lactate refers to squalamine dilactate.
  • squalamine phosphate refers to any phosphate salt of squalamine. In some embodiments, squalamine phosphate refers to squalamine monophosphate. In some embodiments, squalamine phosphate refers to squalamine bis-phosphate. In some embodiments, squalamine phosphate refers to a hydrated phosphate salt of squalamine.
  • the combining step is performed at a temperature of about 55 °C to about 70 °C. This includes a temperature of about 55 °C to about 65 °C, about 55 °C to about 60 °C, about 60 °C to about 70 °C, about 60 °C to about 65 °C, or about 65 °C to about 70 °C.
  • the temperature is about 55, about 56, about 57, about 58, about 59, about 60, about 61 , about 62, about 63, about 64, about 65, about 66, about 67, about 68, about 69, or about 70 °C, including increments therein.
  • the combining step is performed at 60 ⁇ 5 °C.
  • the solution of squalamine lactate is a methanol solution of squalamine lactate.
  • the heating step is performed at a temperature of about 65 °C to about 75 °C. This includes a temperature of about 65, about 66, about 67, about 68, about 69, about 70, about 71 , about 72, about 73, about 74, or about 75 °C, including increments therein. In some embodiments, the heating step is performed at a temperature of about 65 °C to about 70 °C, or about 70 °C to about 75 °C. In some embodiments, the heating step is performed at 70 ⁇ 5 °C. In some embodiments, the heating step is performed at a temperature above the temperature attained during the combining step.
  • the process further comprises stirring the combined mixture after cooling but before the isolating step.
  • the isolating step comprises filtering the combined mixture and washing filtered solids with a solvent. In some embodiments, the isolating step comprises filtering the combined mixture and washing filtered solids with acetone. In some embodiments, the isolating step comprises filtering the combined mixture and washing filtered solids with acetone, water, ethanol, methanol, methyl ethyl ketone, or any combination thereof.
  • a process for preparing the squalamine phosphate solid form of Form 2 comprising: (a) dissolving squalamine lactate in base, water, and alcohol to form a first solution; (b) heating the first solution to a first elevated temperature, wherein the first elevated temperature is greater than 25 °C; (c) adding a first amount of phosphoric acid (H 3 P0 4 ) to the first solution to form a second solution; (d) heating the second solution to a second elevated temperature higher than the first elevated temperature; (e) adding a second amount of H3PO4 to the second solution; (f) obtaining a slurry; and (h) isolating the squalamine phosphate solid form.
  • the base is sodium hydroxide. In some embodiments, the base is potassium hydroxide. In some embodiments, the base is lithium hydroxide.
  • the alcohol is ethanol. In some embodiments, the alcohol is methanol. In some embodiments, the alcohol is propanol. In some embodiments, the alcohol is ethanol, methanol, propanol, or any combination thereof.
  • the first elevated temperature is at least about 35 °C. This includes a first elevated temperature of at least about 36, about 37, about 38, about 39, about 40, about 41 , about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, or about 55 °C, including increments therein. In some embodiments, the first elevated temperature is about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, or about 55 °C, including increments therein.
  • the first elevated temperature is about 35 °C to about 55 °C. This includes the temperature range of about 35 °C to about 50 °C, about 35 °C to about 45 °C, about 35 °C to about 40 °C, about 40 °C to about 55 °C, about 40 °C to about 50 °C, about 40 °C to about 45 °C, about 45 °C to about 55 °C, about 45 °C to about 50 °C, or about 50 °C to about 55 °C.
  • the second elevated temperature is at least about 45 °C. This includes a second elevated temperature of at least about 46, about 47, about 48, about 49, about 50, about 51 , about 52, about 53, about 54, or about 55 °C, including increments therein. In some embodiments, the second elevated temperature is about 45, about 46, about 47, about 48, about 49, about 50, about 51, about 52, about 53, about 54, or about 55 °C, including increments therein. In some embodiments, the second elevated temperature is about 45 °C to about 55 °C. This includes the temperature range of about 45 °C to about 50 °C, or about 50 °C to about 55 °C.
  • the step of obtaining the slurry comprises seeding the mixture.
  • a previous batch of squalamine phosphate is used to seed the mixture, preferably of the same form that is being prepared.
  • the step of obtaining the slurry comprises adding a third amount of H3PO4 to the second solution.
  • the step of obtaining the slurry comprises cooling the second solution. In some embodiments, the step of obtaining the slurry comprises cooling the second solution to room temperature.
  • the process further comprises aging the slurry prior to the isolating step.
  • the step of aging the slurry is performed for at least about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, or about 16 hours.
  • the step of aging the slurry is performed for about 8, about 9, about 10, about 1 1, about 12, about 13, about 14, about 15, or about 16 hours.
  • the isolating step comprises filtering the slurry and washing filtered solids with a solvent. In some embodiments, the isolating step comprises filtering the slurry and washing filtered solids with acetone. In some embodiments, the isolating step comprises filtering the slurry and washing filtered solids with acetone. In some embodiments, the isolating step comprises filtering the slurry and washing filtered solids with acetone, water, ethanol, methanol, methyl ethyl ketone, or any combination thereof.
  • a process for preparing the squalamine phosphate solid form of Form 2 comprising: (a) combining squalamine free base in water and alcohol to form a first solution; (b) adding phosphoric acid (H3PO4) to the first solution to obtain a first slurry; (c) aging the first slurry; (d) isolating solids from the first slurry; (e) mixing the solids in water and alcohol to form a second slurry; (f) heating the second slurry; (g) cooling the second slurry; (h) repeating steps (f) and (g); and (i) isolating the squalamine phosphate solid form of Form 2.
  • the alcohol of step (a) is ethanol.
  • the water and alcohol of step (a) is 90: 10 v/v ethanohwater.
  • the water and alcohol of step (a) is from 90: 10 v/v ethanohwater to 50:50 v/v ethanol: water. This includes 85: 15 v/v ethanol: water, 80:20 v/v ethanohwater, 75:25 v/v ethanohwater, 70:30 v/v
  • ethanohwater 65:35 v/v ethanohwater, 60:40 v/v ethanohwater, 55:45 v/v ethanohwater, and 50:50 v/v ethanohwater, including increments therein.
  • step (a) further comprises heating to form the first solution. In further embodiments, step (a) further comprises heating to 40 °C to form the first solution.
  • step (b) is performed at a lower temperature than step (a). In some embodiments, the lower temperature is about 35 °C. In some embodiments, phosphoric acid is added batch-wise to the first solution. In some embodiments, step (b) further comprises seeding the first solution. In some embodiments, a previous batch of squalamine phosphate is used to seed the first solution, preferably of the same form that is being prepared. [0177] In some embodiments, step (c) is performed at the same temperature as step (b). In some embodiments, step (c) is performed at the same temperature as step (b) for at least about 4 hours and then at a lower temperature for at least about 8 hours.
  • step (c) is performed at the same temperature as step (b) for about 6 hours and then at a lower temperature for about 10 hours.
  • the lower temperature is less than about 35 °C. In still further embodiments, the lower temperature is about 20 °C.
  • step (d) comprises filtering the first slurry and washing filtered solids with a solvent. In some embodiments, step (d) comprises filtering the first slurry and washing filtered solids with acetone.
  • the alcohol of step (e) is ethanol. In some embodiments, the water and alcohol of step (e) is 67:33 v/v ethanol/water.
  • the heating in step (f) is performed at a temperature of at least about 40 °C.
  • the temperature is about 41 , about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 51 , about 52, about 53, about 54, or about 55 °C.
  • the second slurry is cooled to a temperature below about 40 °C in step (g).
  • the temperature is about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, or about 39 °C.
  • steps (f) and (g) are repeated at least about 2, about 3, about 4, about 5, or about 6 times. In some embodiments, steps (f) and (g) are repeated about 2, about 3, about 4, about 5, or about 6 times.
  • step (i) comprises filtering the second slurry and washing filtered solids with a solvent. In some embodiments, step (i) comprises filtering the second slurry and washing filtered solids with acetone.
  • squalamine lactate is replaced with another salt of squalamine, wherein the other salt of squalamine is not squalamine phosphate. In some embodiments, squalamine lactate is converted to another salt of squalamine, wherein the other salt of squalamine is not squalamine phosphate, prior to conversion to squalamine phosphate.
  • substantially generally refers to at least 90% similarity.
  • substantially refers to ⁇ 0.2°.
  • first differential scanning calorimetry thermogram being substantially as shown in a second differential scanning calorimetry thermogram
  • substantially refers to ⁇ 0.4 °C.
  • substantially refers to ⁇ 0.4% weight.
  • substantially purified refers to at least 95% purity. This includes at least 96, 97, 98, or 99% purity. In further embodiments, “substantially purified” refers to about 95, 96, 97, 98, 99, 99.5, or 99.9% purity, including increments therein.
  • therapeutic activity or “activity” may refer to an activity whose effect is consistent with a desirable therapeutic outcome in humans, or to desired effects in non-human mammals or in other species or organisms.
  • Therapeutic activity may be measured in vivo or in vitro. For example, a desirable effect may be assayed in cell culture.
  • the phrase "therapeutically effective amount” shall mean the drug dosage that provides the specific pharmacological response for which the drug is administered in a significant number of subjects in need of such treatment. It is emphasized that a
  • therapeutically effective amount of a drug that is administered to a particular subject in a particular instance will not always be effective in treating the conditions/diseases described herein, even though such dosage is deemed to be a therapeutically effective amount by those of skill in the art.
  • X-ray powder diffraction (XRPD) patterns of the isolated product are shown in FIG. 1 (XRPD patterns (b) and (a) as wet and dry cakes, respectively), indicating Form 2. It is different from the XRPD pattern of Form 1.
  • Squalamine lactate crystals were prepared from the crude Squalamine described in Example 1 via the literature procedure prior to conversion to the lactate salt.
  • a 100-L reactor was equipped with a thermocouple, a nitrogen inlet, a condenser, an addition funnel and a thermal regulation unit.
  • a pre-prepared solution of water (26.5 L), sodium phosphate dibasic (397.0 g, 2.797 mol), and sodium phosphate monobasic (16.0 g, 0.133 mol) was transferred into the 100-L Reactor via a PTFE tubing attached in-line with a 0.2 or 0.4- micron filter cartridge.
  • the contents of the 100 L Reactor were agitated under nitrogen at 25 ⁇ 5 °C at 125-250 RPM, resulting in a clear solution.
  • a pH meter was standardized and then the pH of the phosphate buffer stirring in the 100 L Reactor was determined which was found to be at pH 8.1.
  • Water (13.2 L) and methanol (26.5 L) were added and resulting mixture was heated to 60 ⁇ 5°C over a period of 1.5 to 3.0 h.
  • a solution of squalamine lactate (264.0 g, 0.327 mol) and methanol (1 .32 L) was prepared and transferred into the addition funnel atop the 100 L Reactor via a PTFE tubing attached in-line with a 0.2 or 0.4-micron filter cartridge.
  • the squalamine lactate/MeOH solution in the addition funnel was slowly transferred into the 100 L reactor over a period of 20-25 min while maintaining the temperature at 60 ⁇ 5°C. After the addition was over, the resulting cloudy mixture was heated to 70 ⁇ 5 °C to obtain a near clear solution which contained trace amounts of suspended particles. Heating was continued for at least an hour after which heating was turned off and the batch was allowed to cool to ambient temperature over several hours. The batch was then stirred over night at room temperature. The resulting white suspension was filtered and the cake was dried on the filter funnel until nearly no filtrate could be observed. The resulting white cake was washed with acetone (2 x 5.3 L).
  • Scan from 2 to 40 degrees 2-theta; 0.01 degrees/step; scan rate 2 degrees/min.
  • Form 2 Washing Form 2 with water followed by acetone or ethanol wash and subsequent drying at 40 °C generates Form 1.
  • Form 2 was slurred in dry ethanol at 30 °C it also converted to Form 1.
  • Form 1 was also obtained by treating Form 2 at low humidity (27% RH), while at high humidity (70% RH) Form 2 remained the same.
  • Form 2 is favored at high water content in slurry or high humidity in air, while Form 1 is more stable at lower humidity or low water content.
  • Procedure 3 dry ethanol slurry at 30 °C
  • Form 1 is stable at water activity levels at or lower than 0.5 A w in ethanol.
  • Form 2 will be stable at A w levels 0.6 and above.
  • a blend was prepared by combining 518.6 g of Prosolv SMCC HD 90 (JRS Pharma), 13.4 g of Ac-Di-Sol SD-71 1 (FMC), 1.33 g of Aerosil 200 (Evonik), 3.4 g of magnesium stearate (Malinckrodt), and 135.8 g of squalamine phosphate Form 1 in a V-blender during 10 min to yield 667.6 grams of material (99%).
  • This blend was added to the hopper of a tablet press.
  • the target tablet parameters were a weight of 125 mg, thickness of 4.15 mm, hardness of 12 kp, and friability of NMT 1.0%.
  • the die fill amount and compression parameters were set to achieve tablet requirements (Fill weight of 9.85, turret speed of 20 RPM, main compression of 5.8, and average upper compression of 2.5 N). Using these parameters, 10 sample tablets were prepared that met required properties. The tableting process was initiated and tablets meeting requirements were collected (609.3 g, 91%).
  • the spray gun was loaded with the coating mixture and the squalamine phosphate tablets were loaded in the coating pan at 5.0 inches from the spray gun nozzle.
  • Pre-heat time was set at 6 min 30 sec, spray rate at 4 g/min, pan speed at 15 rpm, inlet temperature at 60-75 °C, exhaust temperature at 45-50 °C, bed temperature at 45-50 °C, atomization at 20 psi, and process air at 150 cfm. Spraying was begun and continued until the theoretical amount of 217.6 gram s of coating suspension was applied.
  • the pan speed was reduced to 3 rpm and the inlet temperature reduced to 0 °C in order to dry the tablets over 10 min.
  • the coated tablets were collected and weighed to yield 626.9 g (99%) of coated squalamine phosphate tablets.

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Abstract

L'invention concerne des formes solides cristallines de phosphate de squalamine désignées par Forme 1 et Forme 2, des compositions contenant une ou les deux formes, et des procédés pour les préparer et les utiliser.
PCT/US2018/057650 2017-10-30 2018-10-26 Nouvelles formes solides de squalamine et procédés pour les produire Ceased WO2019089365A1 (fr)

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JP2023518381A (ja) * 2020-03-19 2023-05-01 エンターイン、 インコーポレイテッド スクアラミンの結晶形

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JP2023518381A (ja) * 2020-03-19 2023-05-01 エンターイン、 インコーポレイテッド スクアラミンの結晶形

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EP3704132A1 (fr) 2020-09-09
US20210332079A1 (en) 2021-10-28

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